Development of a high-throughput assay to detect antibody inhibition of low pH induced conformational changes of influenza virus hemagglutinin

Autor:	Jacqueline M. Katz, Seiji Hongo, Xiuhua Lu, Zhu-Nan Li, Jessica F. Trost, Paul J. Carney, Kanetsu Sugawara, Terrence M. Tumpey, Elizabeth LeMasters, James Stevens, David A. Steinhauer, Feng Liu, Min Z. Levine
Jazyk:	angličtina
Rok vydání:	2018
Předmět:	Models Molecular RNA viruses 0301 basic medicine Conformational change Protein Conformation Physiology Cell Membranes lcsh:Medicine Hemagglutinin Glycoproteins Influenza Virus medicine.disease_cause Biochemistry Membrane Fusion Physical Chemistry law.invention Protein structure law Immune Physiology Medicine and Health Sciences Influenza A virus Enzyme-Linked Immunoassays lcsh:Science Pathology and laboratory medicine Immune System Proteins Multidisciplinary biology Chemistry Proteases Hydrogen-Ion Concentration Medical microbiology Recombinant Proteins Enzymes 3. Good health Physical Sciences Viruses Recombinant DNA Sorption Cellular Structures and Organelles Pathogens Antibody Research Article medicine.drug_class Immunology 030106 microbiology Hemagglutinin (influenza) Enzyme-Linked Immunosorbent Assay Research and Analysis Methods Monoclonal antibody Microbiology Antibodies Virus 03 medical and health sciences Influenza Human Protein Concentration Assays medicine Humans Influenza viruses Immunoassays Molecular Biology Techniques Molecular Biology Molecular Biology Assays and Analysis Techniques Influenza A Virus H3N2 Subtype lcsh:R Organisms Viral pathogens Biology and Life Sciences Proteins Cell Biology Antibodies Neutralizing Molecular biology High-Throughput Screening Assays Microbial pathogens 030104 developmental biology Enzymology Immunologic Techniques biology.protein lcsh:Q Adsorption Serine Proteases Orthomyxoviruses
Zdroj:	PLoS ONE, Vol 13, Iss 6, p e0199683 (2018) PLoS ONE
ISSN:	1932-6203
Popis:	Many broadly neutralizing antibodies (bnAbs) bind to conserved areas of the hemagglutinin (HA) stalk region and can inhibit the low pH induced HA conformational changes necessary for viral membrane fusion activity. We developed and evaluated a high-throughput virus-free and cell-free ELISA based low pH induced HA Conformational Change Inhibition Antibody Detection Assay (HCCIA) and a complementary proteinase susceptibility assay. Human serum samples (n = 150) were tested by HCCIA using H3 recombinant HA. Optical density (OD) ratios of mAb HC31 at pH 4.8 to pH 7.0 ranged from 0.87 to 0.09. Our results demonstrated that low pH induced HA conformational change inhibition antibodies (CCI) neutralized multiple H3 strains after removal of head-binding antibodies. The results suggest that HCCIA can be utilized to detect and characterize CCI in sera, that are potentially broadly neutralizing, and serves as a useful tool for evaluating universal vaccine candidates targeting the HA stalk.
Databáze:	OpenAIRE
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